BRENDA - Enzyme Database show
show all sequences of 1.14.13.24

Molecular and biochemical characterization of 3-hydroxybenzoate 6-hydroxylase from Polaromonas naphthalenivorans CJ2

Park, M.; Jeon, Y.; Jang, H.H.; Ro, H.S.; Park, W.; Madsen, E.L.; Jeon, C.O.; Appl. Environ. Microbiol. 73, 5146-5152 (2007)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
orf2 or gene nagX, which does not encode salicylate 5-hydroxylase, but 3-hydroxybenzoate 6-hydroxylase, DNA and amino acid sequence determination and analysis, genetic organization, overview, overexpression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)
Polaromonas naphthalenivorans
Engineering
Amino acid exchange
Commentary
Organism
additional information
construction of a nagX knockout mutant of strain CJ2 defective in nagX, the mutant fails to grow on 3-hydroxybenzoate but grows normally on naphthalene
Polaromonas naphthalenivorans
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.065
-
3-hydroxybenzoate
pH 8.0, 30C, recombinant NagX, with NADH
Polaromonas naphthalenivorans
0.493
-
NADH
pH 8.0, 30C, recombinant NagX
Polaromonas naphthalenivorans
0.86
-
NADPH
pH 8.0, 30C, recombinant NagX
Polaromonas naphthalenivorans
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
43000
-
recombinant enzyme, gel filtration
Polaromonas naphthalenivorans
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
3-hydroxybenzoate + NAD(P)H + H+ + O2
Polaromonas naphthalenivorans
the small catabolic gene cluster nagR2-orf2I''KL in strain CJ2 may be multifunctional and is essential for the degradation of 3-hydroxybenzoate
gentisate + NAD(P)+ + H2O
i.e. 2,5-dihydroxybenzoate
-
?
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Polaromonas naphthalenivorans
-
orf2, gene nagX
-
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
3-hydroxybenzoate + NAD(P)H + H+ + O2
-
684560
Polaromonas naphthalenivorans
gentisate + NAD(P)+ + H2O
i.e. 2,5-dihydroxybenzoate
-
-
?
3-hydroxybenzoate + NAD(P)H + H+ + O2
the small catabolic gene cluster nagR2-orf2I''KL in strain CJ2 may be multifunctional and is essential for the degradation of 3-hydroxybenzoate
684560
Polaromonas naphthalenivorans
gentisate + NAD(P)+ + H2O
i.e. 2,5-dihydroxybenzoate
-
-
?
additional information
the enzyme shows no salicylate 5-hydroxylase activity
684560
Polaromonas naphthalenivorans
?
-
-
-
-
Subunits
Subunits
Commentary
Organism
monomer
1 * 43000, recombinant enzyme, SDS-PAGE
Polaromonas naphthalenivorans
Temperature Optimum [C]
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
30
-
-
Polaromonas naphthalenivorans
Temperature Range [C]
Temperature Minimum [C]
Temperature Maximum [C]
Commentary
Organism
10
50
inactive above 50C
Polaromonas naphthalenivorans
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
8.4
-
-
Polaromonas naphthalenivorans
pH Range
pH Minimum
pH Maximum
Commentary
Organism
5
9.4
-
Polaromonas naphthalenivorans
Cofactor
Cofactor
Commentary
Organism
Structure
FAD
bound to the enzyme
Polaromonas naphthalenivorans
NADH
-
Polaromonas naphthalenivorans
NADPH
-
Polaromonas naphthalenivorans
Cloned(Commentary) (protein specific)
Commentary
Organism
orf2 or gene nagX, which does not encode salicylate 5-hydroxylase, but 3-hydroxybenzoate 6-hydroxylase, DNA and amino acid sequence determination and analysis, genetic organization, overview, overexpression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)
Polaromonas naphthalenivorans
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
FAD
bound to the enzyme
Polaromonas naphthalenivorans
NADH
-
Polaromonas naphthalenivorans
NADPH
-
Polaromonas naphthalenivorans
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
additional information
construction of a nagX knockout mutant of strain CJ2 defective in nagX, the mutant fails to grow on 3-hydroxybenzoate but grows normally on naphthalene
Polaromonas naphthalenivorans
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.065
-
3-hydroxybenzoate
pH 8.0, 30C, recombinant NagX, with NADH
Polaromonas naphthalenivorans
0.493
-
NADH
pH 8.0, 30C, recombinant NagX
Polaromonas naphthalenivorans
0.86
-
NADPH
pH 8.0, 30C, recombinant NagX
Polaromonas naphthalenivorans
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
43000
-
recombinant enzyme, gel filtration
Polaromonas naphthalenivorans
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
3-hydroxybenzoate + NAD(P)H + H+ + O2
Polaromonas naphthalenivorans
the small catabolic gene cluster nagR2-orf2I''KL in strain CJ2 may be multifunctional and is essential for the degradation of 3-hydroxybenzoate
gentisate + NAD(P)+ + H2O
i.e. 2,5-dihydroxybenzoate
-
?
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
3-hydroxybenzoate + NAD(P)H + H+ + O2
-
684560
Polaromonas naphthalenivorans
gentisate + NAD(P)+ + H2O
i.e. 2,5-dihydroxybenzoate
-
-
?
3-hydroxybenzoate + NAD(P)H + H+ + O2
the small catabolic gene cluster nagR2-orf2I''KL in strain CJ2 may be multifunctional and is essential for the degradation of 3-hydroxybenzoate
684560
Polaromonas naphthalenivorans
gentisate + NAD(P)+ + H2O
i.e. 2,5-dihydroxybenzoate
-
-
?
additional information
the enzyme shows no salicylate 5-hydroxylase activity
684560
Polaromonas naphthalenivorans
?
-
-
-
-
Subunits (protein specific)
Subunits
Commentary
Organism
monomer
1 * 43000, recombinant enzyme, SDS-PAGE
Polaromonas naphthalenivorans
Temperature Optimum [C] (protein specific)
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
30
-
-
Polaromonas naphthalenivorans
Temperature Range [C] (protein specific)
Temperature Minimum [C]
Temperature Maximum [C]
Commentary
Organism
10
50
inactive above 50C
Polaromonas naphthalenivorans
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
8.4
-
-
Polaromonas naphthalenivorans
pH Range (protein specific)
pH Minimum
pH Maximum
Commentary
Organism
5
9.4
-
Polaromonas naphthalenivorans
Other publictions for EC 1.14.13.24
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [C]
Temperature Range [C]
Temperature Stability [C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [C] (protein specific)
Temperature Range [C] (protein specific)
Temperature Stability [C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
742582
Montersino
3-Hydroxybenzoate 6-hydroxyla ...
Rhodococcus jostii
Front. Microbiol.
8
1110
2017
-
-
1
-
-
-
-
-
-
-
-
-
-
5
-
-
1
-
-
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2
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1
2
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1
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-
-
-
-
-
-
-
1
1
-
-
-
742501
Sucharitakul
Tyr217 and His213 are importa ...
Rhodococcus jostii
FEBS J.
283
860-881
2016
-
-
-
-
7
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
1
-
7
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
743771
Srisook
-
Immobilization of 3-hydroxybe ...
Rhodococcus jostii
React. Funct. Polym.
82
41-46
2014
-
-
-
-
-
1
-
3
-
-
-
-
-
1
-
-
-
-
-
-
-
-
1
-
-
-
-
2
2
2
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
3
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
2
2
2
-
-
-
-
-
-
-
-
727958
Montersino
Crystal structure of 3-hydroxy ...
Rhodococcus jostii
J. Biol. Chem.
288
26235-26245
2013
-
-
1
1
4
-
-
2
-
-
-
1
-
3
-
-
-
-
-
-
-
-
1
1
-
-
-
2
-
-
-
2
-
-
-
-
-
1
2
1
4
-
-
-
-
2
-
-
-
1
-
-
-
-
-
-
-
-
1
1
-
-
-
2
-
-
-
-
-
-
-
-
-
-
727968
Sucharitakul
The reaction kinetics of 3-hyd ...
Rhodococcus jostii
J. Biol. Chem.
288
35210-35221
2013
-
-
-
-
-
-
-
-
-
-
-
1
-
2
-
-
-
-
-
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-
1
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-
-
1
-
-
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2
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2
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1
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-
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-
1
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
726983
Sucharitakul
Reduction kinetics of 3-hydrox ...
Rhodococcus jostii
Biochemistry
51
4309-4321
2012
-
-
1
-
-
-
-
-
-
-
1
1
-
2
-
-
1
-
-
-
-
-
1
1
-
-
-
-
-
-
-
2
-
-
-
-
-
1
2
-
-
-
-
-
-
-
-
-
1
1
-
-
-
1
-
-
-
-
1
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
727077
Montersino
Functional annotation and char ...
Rhodococcus jostii
Biochim. Biophys. Acta
1824
433-442
2012
3
-
1
-
-
-
1
6
-
-
2
1
-
5
-
-
1
-
-
-
2
-
10
1
-
-
-
4
1
-
-
2
-
-
-
3
-
1
2
-
-
-
-
1
-
6
-
-
2
1
-
-
-
1
-
-
2
-
10
1
-
-
-
4
1
-
-
-
-
-
-
-
4
4
728255
Holesova
Gentisate and 3-oxoadipate pat ...
Candida parapsilosis, Candida parapsilosis CDU1
Microbiology
157
2152-2163
2011
-
-
-
-
-
-
-
-
1
-
1
2
-
4
-
-
-
-
-
-
-
-
2
1
-
-
-
-
-
-
-
2
-
-
-
-
-
-
2
-
-
-
-
-
-
-
1
-
1
2
-
-
-
-
-
-
-
-
2
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
710986
Liu
Functional characterization of ...
Rhodococcus sp., Rhodococcus sp. NCIMB 12038
Appl. Microbiol. Biotechnol.
90
671-678
2010
-
-
1
-
-
-
-
-
-
-
2
4
-
3
-
-
1
-
-
1
1
-
8
1
1
-
-
-
1
-
-
3
-
-
-
-
-
1
3
-
-
-
-
-
-
-
-
-
2
4
-
-
-
1
-
1
1
-
8
1
1
-
-
-
1
-
-
-
-
2
2
-
-
-
712313
Yang
Purification and characterizat ...
Corynebacterium glutamicum, Corynebacterium glutamicum ATCC 13032
J. Basic Microbiol.
50
599-604
2010
-
-
1
-
-
-
-
1
-
-
2
4
-
4
-
-
1
1
-
-
1
-
4
1
1
-
-
-
1
1
-
3
-
-
-
-
-
1
3
-
-
-
-
-
-
1
-
-
2
4
-
-
-
1
-
-
1
-
4
1
1
-
-
-
1
1
-
-
-
2
2
-
-
-
684560
Park
Molecular and biochemical char ...
Polaromonas naphthalenivorans
Appl. Environ. Microbiol.
73
5146-5152
2007
-
-
1
-
1
-
-
3
-
-
1
1
-
5
-
-
-
-
-
-
-
-
3
1
1
1
-
-
1
1
-
3
-
-
-
-
-
1
3
-
1
-
-
-
-
3
-
-
1
1
-
-
-
-
-
-
-
-
3
1
1
1
-
-
1
1
-
-
-
-
-
-
-
-
659871
Liu
Arg169 is essential for cataly ...
Klebsiella oxytoca
Microbiol. Res.
160
53-59
2005
-
-
1
-
1
-
-
-
-
-
1
1
-
2
-
-
-
1
-
-
1
-
1
2
1
-
-
-
1
-
-
2
-
-
-
-
-
1
2
-
1
-
-
-
-
-
-
-
1
1
-
-
-
-
-
-
1
-
1
2
1
-
-
-
1
-
-
-
-
-
-
-
-
-
674255
Gao
Molecular and biochemical char ...
Pseudomonas alcaligenes, Pseudomonas alcaligenes P25X
J. Bacteriol.
187
7696-7702
2005
1
-
1
-
1
-
6
4
-
1
2
4
-
4
-
-
1
-
-
1
1
3
16
1
1
1
-
-
1
1
-
2
-
-
-
1
-
1
2
-
1
-
-
6
-
4
-
1
2
4
-
-
-
1
-
1
1
3
16
1
1
1
-
-
1
1
-
-
-
-
-
-
-
-
438962
Suresh
Interaction of 3-hydroxybenzoa ...
Micrococcus sp.
Biotechnol. Prog.
17
1026-1031
2001
-
-
-
-
-
-
4
1
-
-
-
-
-
1
-
-
1
-
-
-
-
-
1
-
-
-
-
-
1
-
-
1
-
-
-
-
-
-
1
-
-
-
-
4
-
1
-
-
-
-
-
-
-
1
-
-
-
-
1
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
438961
Sumathi
Reactivity of 3-HBA-6-hydroxyl ...
Micrococcus sp.
Biotechnol. Prog.
16
577-582
2000
-
-
-
-
-
-
2
-
-
-
-
-
-
1
-
-
1
-
-
-
-
-
1
-
-
-
-
-
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2
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1
-
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-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
438960
Sumathi
Chemical modification of 3-HBA ...
Micrococcus sp.
Indian J. Biochem. Biophys.
35
266-272
1998
-
-
-
-
-
-
1
-
-
-
-
-
-
2
-
-
1
-
-
-
-
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-
-
1
-
-
-
-
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-
-
-
-
-
-
-
-
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-
-
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1
-
-
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1
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
438958
Suarez
Purification and characterizat ...
Klebsiella pneumoniae, Klebsiella pneumoniae M5a1
FEMS Microbiol. Lett.
126
283-290
1995
-
-
-
-
-
1
3
2
-
-
2
-
-
12
-
-
1
-
-
-
1
1
2
1
1
1
-
-
1
1
1
2
-
-
-
-
-
-
2
-
-
1
-
3
-
2
-
-
2
-
-
-
-
1
-
-
1
1
2
1
1
1
-
-
1
1
1
-
-
-
-
-
-
-
438959
Suemori
-
Physicochemical and immunochem ...
Rhodococcus erythropolis, Rhodococcus erythropolis S1
Biotechnol. Lett.
17
1063-1068
1995
-
-
-
-
-
-
-
-
-
-
-
-
-
5
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
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-
-
1
-
-
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-
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-
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Pseudomonas cepacia 3-hydroxyb ...
Burkholderia cepacia
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1987
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Pseudomonas cepacia 3-hydroxyb ...
Burkholderia cepacia
Biochemistry
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1105-1110
1987
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The stereospecificity of bacte ...
Pseudomonas aeruginosa, Pseudomonas aeruginosa T1
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1977
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438953
Groseclose
3-Hydroxybenzoate 6-hydroxylas ...
Pseudomonas aeruginosa, Pseudomonas aeruginosa T1
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897-903
1973
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